The analysis of hepatitis B virus (HBV) mutations is important for understanding HBV progression and for deciding on appropriate clinical treatments. However, it is difficult to determine the quantitative abundance of various mutants in heterogeneous mixtures by conventional methods such as direct sequencing or the TaqMan assay. In this study, we investigated the possibility of using both allele-specific oligonucleotide hybridization (ASOH) and allele-specific oligonucleotide competitive hybridization (ASOCH) with the Handy Bio-Strand system for the quantitative identification of three well-defined HBV variants: the basal core promoter (BCP) mutations (nt1762 and nt1764), the pre-core (PC) mutation (nt1896), and variance at nt1858. Using standardized mixtures of wild-type and mutant DNA, optimal hybridization conditions for ASOH and ASOCH were determined. Next, the performance of these methods was evaluated using actual serum DNAs from HBV patients. Excellent reproducibility was obtained both in the analysis of internal positive controls and in the semi-quantitative categorization of heterogeneous viral mixtures into five abundance groups (0%, 25%, 50%, 75%, and 100% mutant virus). Combined with real-time PCR to determine the HBV viral load, this hybridization method offers a new tool with applications both in HBV clinical research and treatment.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.jbiosc.2009.06.023 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Tandem repeat disorders: from diagnosis to emerging therapeutic strategies.
Encephalitis
December 2024
Rare Disease Center, Department of Genomic Medicine; Department of Neurology, Seoul National University Hospital, Seoul National University College of Medicine, Seoul, Korea.
Article Synopsis
- * These disorders present unique clinical features, such as reduced penetrance and changes in repeat motifs, and advancements in molecular diagnostics, especially long-read sequencing, have improved detection rates for TRDs.
- * Innovative therapeutic approaches, including antisense oligonucleotides (ASOs) and gene editing technologies, show promise for treating these conditions, but further research is needed to enhance treatment efficacy and patient outcomes.
A chip-based universal strategy to realize multiplex PCR by using wax films for sealing and controllable release of primers.
Biosens Bioelectron
February 2025
School of Biomedical Engineering, Tsinghua University, Beijing, 100084, China; National Engineering Research Center for Beijing Biochip Technology, Beijing, 102200, China. Electronic address:
Simultaneous detection of multiple nucleic acid targets from a single sample is a common requirement in molecular diagnosis and basic research. Dividing a bulky polymerase chain reaction (PCR) into many isolated small reaction units through microfluidic technology is commonly used to realize this goal. However, previous microfluidic platforms for multiplex PCR suffer from complex structures and strict operation requirements.
View Article and Find Full Text PDFHigh-Resolution Melting Assay to Detect the Mutations That Cause the Y132F and G458S Substitutions at the ERG11 Gene Involved in Azole Resistance in Candida parapsilosis.
Mycoses
November 2024
Mycology Reference Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain.
Background: Candida parapsilosis is a pathogenic yeast that has reduced susceptibility to echinocandins and ranks as the second or third leading cause of candidaemia, depending on the geographical region. This yeast often causes nosocomial infections, which are frequently detected as outbreaks. In recent years, resistance to azoles in C.
View Article and Find Full Text PDFAllele-specific silencing of a dominant mutation in familial amyotrophic lateral sclerosis type 4.
bioRxiv
October 2024
National Institute of Neurological Disorders and Stroke, National Institutes of Health, 35 Convent Dr., Bethesda, MD 20892, USA.
Amyotrophic lateral sclerosis 4 (ALS4) is an autosomal dominant motor neuron disease that is molecularly characterized by reduced R-loop levels and caused by pathogenic variants in (). encodes an RNA/DNA helicase that resolves three-stranded nucleic acid structures called R-loops. Currently, there are no disease-modifying therapies available for ALS4.
View Article and Find Full Text PDFFunctional characterization of QT interval associated SCN5A enhancer variants identify combined additive effects.
HGG Adv
January 2025
Institute of Molecular Medicine, McGovern Medical School, University of Texas Health Science Center at Houston, Houston, TX 77030, USA. Electronic address:
Several empirical and theoretical studies suggest the presence of multiple enhancers per gene that collectively regulate gene expression, and that common sequence variation impacting on the activities of these enhancers is a major source of inter-individual gene expression variability. However, for the vast majority of genes, enhancers and the underlying regulatory variation remains unknown. Even for the genes with well-characterized enhancers, the nature of the combined effects from multiple enhancers and their variants, when known, on gene expression regulation remains unexplored.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!